KR900001076B1 - Process for the preparation of n-(4-(3-aminopropyl) aminobutyl)-2,2-dihydroxy ethaneamide - Google Patents

Abstract

N-[4-(3-aminopropyl) aminobutyl -2,2-dihydroxy ethaneamide of formula (II) is prepared by oxidative splitting of the C-C bond of the cpd. of formula (I). In the formula, R1 is either hydroxy gp. or amino gp. (provided that both R1 gps are not amino gp.), and R2 is H, carboxyl or CONH(CH2)4NH2(CH2)3NH2. The cpd. (I), or glyoxylspermidine, has an imuno-modifying effect and is used in the synthesis of spergualin analogues, which are used to treat cancer.

Description

Method for preparing N- [4- (3-aminopropyl) aminobutyl] -2,2-dihydroxyethinamide

The present invention has an immunomodulatory effect on its own and is useful as an intermediate for the synthesis of the anti-cancer substance, spergualin related compounds, N- [4- (3-aminopropyl) aminobutyl represented by the following structural formula (II) ] -2,2-dihydroxy ethaneamide (henceforth a "glyoxylylspermidine").

The present invention also uses N- [4- (3-aminopropyl) aminobutyl] -2- (W-guanidino fatty acid amide)-represented by the following general formula (XV) using the above-described glyoxylylspermidine. The present invention relates to a method for producing a 2-hydroxyethanamide (hereinafter referred to as "15-deoxyspergualin related compound") anticancer substance.

(n is an integer from 6 to 8)

The present inventors have previously used the hydrolysis of a spetualine anticancer substance obtained from culture broth of the bacterium Bacillus laterosporus BMG162-aF ₂ (deposited to No. 5230 in a fermentation research institution). A method for preparing glyoxyyl spermidine has been proposed [Japanese Patent Laid-Open No. 52,263 / 1984; Antibiotic Publication 34, 1622 (1981)].

The present inventors have also proposed a synthesis method using 3-amino-1-propanol as one of the starting materials [Japanese Patent Publication No. 192,347 / 1982; Antibiotic Publication No. 34, 1625 (1981).

The above methods are excellent methods for producing glyoxyyl spermidine, but there are some problems as methods for producing large quantities at low industrial cost.

It is difficult to obtain a large amount of the target product because the first method of purifying and isolating spergualin from a microbial culture using a natural material of severguline as a raw material is very demanding.

In addition, the second method, which is a synthesis method using 3-amino-1-propanol as a starting material, must not only use glyoxylic acid, which is another raw material represented by the following structural formula (III), which is difficult to obtain, The tricky aldehyde-protecting group must be introduced and removed.

Examples of the aldehyde-protecting derivatives include acetals, thioacetals, hydrazones, oximes and diacyl derivatives.

The above-described conventional synthesis methods that go through the step of protecting the aldehyde group of glyoxylic acid have a problem as a large production method because the process is difficult.

It is a first object of the present invention to provide a simple synthesis of glyoxyylspermidine, and a second object of the present invention is to use the aforementioned glyoxyylspermidine to It is to provide a manufacturing method.

More briefly, the present invention provides a method for producing glyoxyyl spermidine represented by the following structural formula (II), characterized by selectively oxidatively cleaving an appropriate CC bond of a compound represented by the following general formula (I). It is about.

In the above formula, R ¹ is a hydroxy group or an amino group (wherein R ¹ may not all be an amino group), and R ² is a hydrogen atom, a substitutable alkyl group, a carboxyl group, a substitutable alkoxycarbonyl group or a carbamoyl group.

The present inventors conducted extensive research to solve the problems of the conventional methods and found a new simple synthesis method that does not require the protection and deprotection operation of the aldehyde group.

According to the reaction mechanism according to the present invention, oxidative cleavage of the compound represented by the general formula (I) as a starting material irrespective of the substituent R ² gives a compound of the formula (II). This oxidation reaction itself oxidizes and cleaves the CC bonds of adjacent diols or their equivalent functional groups to obtain an aldehyde or ketone group. As a known reaction, commercially available reagents or methods can be applied. Periodic acid is mentioned as a reaction reagent which can be used extensively with respect to the compound of general formula (I). In the case where all of R ¹ in General Formula (I) are hydroxy groups, oxygen in the presence of a lead tetraacetate, an iodosil compound, an organic acid cobalt (II) salt catalyst, a sulfuric acid peroxide (I) salt, and thallium nitrate (III) A suitable reaction reagent selected from the group consisting of, ethoxytalum (I), cerium sulfate (IV), bisulfate and nickel peroxide can be used with periodic acid.

As long as the reaction proceeds ideally, the reaction conditions may vary depending on the measurement reagent used. For example, water, acetic acid, trichloro acetic acid, methanol, ethanol, ether, dioxane, benzene, benzonitrile, acetonitrile, pyridine, 4-cyanopyridine, N, N-dimethylformamide, anisole, as reaction solvents, Chlorobenzene, sulfolane, etc. can be used individually or in combination. The reaction temperature can also vary widely from 0 ° C. to the boiling point of the solvent, and the reaction time also varies depending on the specific reaction conditions, generally requiring several minutes to several days.

As a reagent for oxidative fission of the compound represented by the above general formula (I), which is a water-soluble compound, periodic acid is particularly preferable among the above-described reaction reagents. For example, ortho periodate (H ₅ IO ₆ ), sodium meta periodate ( NalO ₄ ) or potassium meta periodate (KlO ₄ ) is preferred. In the case of using these periodic acids, the reaction can generally be carried out in an aqueous solution, but may be carried out in an aqueous solution containing organic solvents such as alcohols (eg, methanol, ethanol), dioxane, ether and the like.

The compound represented by the general formula (I), which is the starting compound of the present reaction, condenses the corresponding acid component having a functional group to protect, if necessary, with an amine component by a known reaction to form an amide bond, and then, if necessary, It can synthesize | combine by deprotection. Acids having a hydroxyl group or an amino group at the α- and β-positions which can be used as starting materials can be easily obtained at low cost from various natural and synthetic compounds. For example, β-hydroxy-α-amino acids (e.g. serine, threonine), polyhydroxy monocarboxylic acids (e.g. glycerin acid, various aldonic acids) and dihydroxydicarboxylic acids ( Examples: tartaric acid, α, β-dihydroxyglutaric acid) can be used.

(X¹및 X²는 각각 아미노-보호기임), 다음 구조식(V)을 갖는 N-(2-시아노에틸)-1,4-디아미노부탄(이하, ＂시아노에틸푸트레신＂이라한다.), H₂N(CH₂)₄NH(CH₂)₂CN(V) 및 다음 구조식(VI)을 갖는 1,4-디아미노부탄(이하 ＂푸트레신＂이라 한다.) H₂N(CH₂)₄NH₂(VI)등을 사용할 수 있다.As an amine raw material, spermidine having a protected functional group or a precursor for synthesis of spermidine can be used if necessary. For example, 1,5,10-triazadecan, wherein both N ¹ and N ⁵ atoms represented by the following general formula (VI) are protected,

(X ¹ and X ² are each an amino-protecting group), N- (2-cyanoethyl) -1,4-diaminobutane having the following structural formula (V) _{and.), H 2 N (CH} 2) 4 NH (CH 2) 2 CN (V) and the following is referred to as 1,4-diaminobutane (hereinafter referred to as "new Trail Fu" having the structural formula (VI).) H ₂ N (CH ₂ ) ₄ NH ₂ (VI) and the like can be used.

Examples of synthesizing the compound represented by Formula (I) by reacting the aforementioned acid components with the amine component will be described later again.

를 수득한 다음, 아미노-보호기를 제거한 후 니트릴을 아미노메틸기로 환원시키거나, 또는 니트릴기를 아미노메틸기로 환원시킨 후 아미노보호기를 제거시키면 목적 화합물(이하, ＂N-세릴-스페르미딘＂이라한다)이 수득된다.In the formula (I), R ¹ is an amino and a hydroxy group using serine or other suitable amino acid (e.g. threonine) as an acid component and cyano ethylputrescine represented by the structural formula (V) as an amino component, and R ² is The compound which is a hydrogen atom or an alkyl group (for example, methyl group) can be synthesized. Moreover, the compound whose R <^2> is a hydrogen atom in general formula (I) can be obtained as follows; Serine is first protected by a known amino-protecting group (e.g., benzyloxycarbonyl group (Z group)), and then condensed with cyano ethylputrescine of formula (I) according to a known amide bond formation reaction. Compound of (iii),

Is obtained, and then the nitrile is reduced to the aminomethyl group after removing the amino-protecting group, or the amino protecting group is removed after reducing the nitrile group to the aminomethyl group, referred to as "N-seryl-spermidine". ) Is obtained.

In general formula (I), the compound whose R <^2> is a alkyl group (for example, methyl group) can be synthesize | combined by the method similar to the above using threonine as a raw material.

을 구조식(VI)의 푸트레신과 반응시킨 다음, 아세틸기를 알카리-가수분해시켜 다음 구조식(IX)의 N-(4-아미노부틸)타르타르산 모노아미드

을 수득하고, 화합물(IX)에 다음 구조식(X)의 아크릴로니트릴, CH₂=CHCN(X)을 작용시켜 N-시아노에틸화하여 다음 구조식(XI)의 N-[4-(2-시아노에틸)아미노부틸]타르타르산 모노아미드,

를 수득한 다음, 화합물(XI)의 니트릴기를 아미노메틸기로 환원시키면 목적 화합물이 수득된다.In general formula (I), the compound in which R <^1> is all a hydroxyl group and R <^2> is a carboxy group can be obtained as follows: Diacetyl tartaric anhydride of following formula (VIII),

Is reacted with putrescine of formula (VI), and then the acetyl group is alkali-hydrolyzed to form N- (4-aminobutyl) tartaric acid monoamide of formula (IX)

Was obtained, and the compound (IX) was subjected to N-cyanoethylation with acrylonitrile of the following formula (X), CH ₂ = CHCN (X) to give N- [4- (2- Cyanoethyl) aminobutyl] tartaric acid monoamide,

Was obtained, and then the nitrile group of compound (XI) was reduced to aminomethyl group to give the desired compound.

In general formula (I), a compound in which R ¹ is all a hydroxy group and R ² is an alkoxycarbonyl group is subjected to ordinary esterification of the carboxy group of compound (I) obtained by the above method, or compound (XI). The carboxy group of) can be obtained by carrying out the usual esterification followed by reduction of the nitrile group. Moreover, in General Formula (I), the compound whose R <^1> is all a hydroxyl group and R <^2> is a carbamoyl group can be synthesize | combined by amino-decomposing the compound (R <^2> alkoxycarbonyl group) obtained by the above-mentioned method.

를 구조식(V)의 시아노에틸푸트레신과 반응시켜 다음 구조식(XIII)의 N,N'-비스[4-(2-시아노에틸)아미노부틸]타르타르산 아미드,

를 수득한 다음, 화합물(XIII)의 니트릴기를 아미노 메틸기로 환원시키면 목적 화합물이 수득된다.In particular, and R ¹ are both hydroxy in the formula (I), R ² is _{-CONH (CH 2) 4 NH (} CH 2) 3 NH 2 group compounds can be synthesized in good yield by the following method: tartaric acid Diethyl ester of tartaric acid of formula (XII), which is readily obtained by known esterification of

To N, N'-bis [4- (2-cyanoethyl) aminobutyl] tartaric acid amide of formula (XIII),

Was obtained, and then the nitrile group of compound (XIII) was reduced to amino methyl group to give the desired compound.

The method of the present invention does not require an aldehyde group-protection operation, as described in detail above, or an aldehyde group protection operation for reducing the glyoxyylspermidine to the original aldehyde group in the final step. Not only can greatly shorten the manufacturing process, but also obtain the desired compound in good yield, and the raw materials for carrying out the present invention are easily available and inexpensive from various natural and synthetic compounds. Has an advantage.

Accordingly, the present invention provides a method for producing glyoxyl spermidine at low cost.

The glyoxyylspermidine prepared above can be used to produce 15-deoxy spergualline related compounds more economically and advantageously than before.

(여기에서, n은 6내지 8의 정수임)와 축합시키면 다음 일반식(XV)의 15-데옥시 스페르구알린 관련 화합물,

(여기에서, n은 상기에서 정의한 바와 같이)이 제조된다.Accordingly, according to the present invention glyoxylylspermidine of formula (II) is substituted with W-guanidino fatty acid amide of formula (XVI),

(Where n is an integer from 6 to 8), condensed with 15-deoxy spergualin related compounds of formula (XV),

Where n is as defined above.

The condensation reaction of glyoxyyl spermidine of formula (II) with W-guanidino fatty acid amide of formula (XVI) can be carried out by the method described in detail in Japanese Patent Application Laid-Open No. 83-62152. . In other words, the reaction is preferably carried out in the same solvent containing no water, but the compounds of formulas (II) and (XVI) are usually in the form of acid addition salts, so they are carried out in the presence of a small amount of water in consideration of solubility. . The amount of water present should be as small as possible as long as the compounds of the formulas (II) and (XVI) are uniformly dissolved. The amount of water is 4 to 40 moles with respect to 1 mole of the compound of the formula (II). Compounds of formulas (II) and (XVI) are usually in acid addition salt form and do not need to be added with acid. However, it is preferable to use an acid catalyst in consideration of the yield. Suitable acid catalysts include inorganic livers (e.g. hydrochloric acid, sulfuric acid, boric acid, etc.) and organic acids (e.g. acetic acid, citric acid, tartaric acid, succinic acid, glutaric acid, adipic acid, etc.), among them carboxyl such as citric acid and glutaric acid. Acids are preferred. The amount of acid used is 0 to 10 moles, preferably 0.5 to 4 moles per one mole of the compound of formula II. Reaction temperature is room temperature to 80 degreeC, and 40-60 degreeC is preferable. The reaction time depends on the specific reaction temperature, and several hours to several days are preferable for obtaining a high yield.

The present invention will be described in more detail with reference to the following Examples, which are not intended to limit the present invention.

Example 1

1. N ¹ - (N'- carbonyldiimidazole benzyloxycarbonyl -L- Sheryl) -N ² - Synthesis of (2-cyanoethyl) -1,4-diaminobutane (VII)

47.8 g (0.2 mol) of N-carbenzyloxy-L-serine was dissolved in 200 ml of dioxane, and 25.3 g (0.22 mol) of N-hydroxysuccinimide was added thereto, followed by dicyclohexyl carbodiimide (DCC) 50 ml of a dioxane solution containing 45.4 g (0.22 mol) was added dropwise with ice cooling with stirring.

After stirring overnight at room temperature, the precipitated dicyclohexylurea was filtered, and the filtrate was concentrated under reduced pressure. The residual solid produced above was dissolved in 150 ml of ethyl acetate, and the resulting solution was added dropwise to 300 ml of ethyl acetate solution containing 42.36 g (0.3 mole) of cyano ethyl putrescine (V). Then, 300 ml of ethyl acetate was added to the reaction solution, and after stirring overnight, 100 ml of ethyl acetate, 100 ml of saturated aqueous sodium bicarbonate solution and 50 ml of water were added, followed by stirring for 1 hour.

The reaction solution was separated into an organic layer and an aqueous layer, and the organic layer was washed with 200 ml of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The remaining solid was dissolved in 50 ml of ethanol, the resulting solution was acidified with concentrated hydrochloric acid while ice-cooled, left at 5 ° C. overnight, and the precipitated product was filtered to give N ^2- (N'-carbenzyloxy-L - Sheryl) -N ² - (2-cyanoethyl) -1,4-diaminobutane (VII) · mono-hydrochloride 26.7g of white crystals were obtained. The aqueous layer was extracted several times with 200 ml of ethyl acetate, the ethyl acetate layer was washed with 200 ml of saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate and then concentrated under reduced pressure. The resulting solid was dissolved in 65 ml of ethanol, the solution was acidified with concentrated hydrochloric acid while ice-cooled and left overnight at 5 ° C. (VII) to give 34.5 g of monohydrochloride. The total yield of (VII). Monohydrochloride was 61.2 g (yield, 76.7%). Melting point 147-149 ° C.

NMR (CD ₃ OD): δ 1.4-1.75 (CH ₂ X ₂ ), 2.4-3.0 (CH ₂ N X ₂ , CH ₂ CN), 3.1-3.4 (CONHCH ₂ ), 3.7 (CH ₂ OH, d), 4.1- 4.3 (CHNH), 5.07

IR (KBr): υ (cm ^-1 ) 3290, 3060, 2940, 2870, 2240, 1700, 1640, 1540, 1475, 1365, 1300, 1240, 1140, 1100, 1020, 700

2. N ¹ -L- Sheryl -N ² - Synthesis of (2-cyanoethyl) -1,4-diaminobutane (XIV)

In methanol 400ml N ² - (N'- carbonyldiimidazole benzyloxycarbonyl -L- Sheryl) -N ² - (2- cyanoethyl) -1,4-dia Dino butane (XII) mono hydrochloride 20g (0.05 mol) After heat dissolution, 1 g of 10% palladium carbon was added, and hydrogenolysis was carried out while passing hydrogen gas at 40 ° C. for 3 hours.

50W x8(다우 케미칼 회사제품, H⁺형)250ml가 충전된 칼럼에 통과시킨 다음 물 1l로 세척한 후 2N 암모니아 수용액으로 용출시켰다. 유효 획분들을 모으고 감압농축시켜 시럽상의 N¹-세릴-N²-(2-시아노에틸)-1,4-디아미노부탄(XIV) 9.18g(수율,80.4%)을 수득한다.After the reaction was completed, nitrogen gas was passed for 30 minutes and the catalyst was filtered off. The filtrate was concentrated under reduced pressure, and the remaining solid was dissolved in 100 ml of water.

250 ml of 50 W x8 (Dow Chemical Co., Ltd., H ⁺ type) was passed through a packed column, washed with 1 L of water, and eluted with a 2N aqueous ammonia solution. Concentrated under reduced pressure to collect the effective stroke by those N ¹ on the syrup - to give the (2-cyanoethyl) -1,4-diaminobutane (XIV) 9.18g (yield, 80.4%) Sheryl -N ^2.

NMR (CD ₃ OD): δ 1.4-1.8 (CH ₂ X ₂ ), 2.5-3.05 (CH ₂ N X ₂ , CH ₂ CN), 3.2-3.5 (CONHCH ₂ , CHNH ₂ ), 3.65 (CH ₂ OH, d)

IR (KBr): υ (cm ^-1 ) 3280, 3060, 2930, 2850, 2230, 1650, 1540, 1460, 1360, 1265, 1120, 1050

3. Synthesis of N-seryl-spermidine (I)

(Pharmacia Co. 제품) 2l가 충전된 칼럼에 통과시킨 다음, 물 2l로 세척한 후 물 3l와 1 몰의 NaCl 수용액 3l로 경사용출을 행하였다. 유효 획분들을 모으고 감압건고시켰다. 잔류하는 고상물을 메탄올로 추출을 행하여 메탄올 추출액을 세파덱스

LH-20 500ml가 충전된 칼럼에 통과시키고 메탄올로 용출시켰다. 유효 획분들을 모으고 감압농축시켜 시럽상의 N-세릴-스페르미딘(I)·트리히드로 클로라이드 5.759g(수율,43.63%)을 수득하였다.In methanol 200ml N ¹ - Sheryl -N ² - (2- cyanoethyl) -1,4-diaminobutane (XIV) 8.82g (36.6 milli moles) was dissolved, here CoCl _2, 6H ₂ O 11.04g (46.4 mmol) and then 8.77 g (231.8 mmol) of NaBH ₄ were added slowly with ice cooling. After the addition was completed, the reaction solution was stirred at room temperature for 2 hours, 200 ml of water was added thereto, and then 2N hydrochloric acid was added to adjust the pH to 6.0. The black precipitate was filtered off and the filtrate was concentrated under reduced pressure. The resulting solids are dissolved in 200 ml of water and CM-Sepadex

After passing through a column filled with 2 l of Pharmacia Co., the mixture was washed with 2 l of water and then eluted with 3 l of water and 3 l of 1 mol of NaCl aqueous solution. Effective fractions were collected and dried under reduced pressure. The remaining solids are extracted with methanol, and the methanol extract is separated by Sephadex.

500 ml of LH-20 was passed through a packed column and eluted with methanol. The effective fractions were collected and concentrated under reduced pressure to yield 5.759 g (yield, 43.63%) of N-seryl-spermidine (I) trihydrochloride on syrup.

NMR (CD ₃ OD): δ 1.5-2.5 (CH ₂ X 3), 2.9-3.5 (CH ₂ N X 4), 3.95 (CH ₂ OH), 4.1 (CHNH ₂ )

IR (KBr): υ (cm ^-1 ) 3410, 3030, 1670, 1620, 1560, 1460, 1270, 1160, 1060

4. Synthesis of glyoxyyl spermidine (II)

500ml가 충전된 칼럼에 통과시켰다. 다음에 물 2.5l와 1M NaCl수용액 2.5l를 사용하여 경사 용출을 수행하여 유효 획분들을 모으고 감압 건고시켰다. 얻어지는 잔류 고상물을 메탄올로 추출을 행하고, 메탄올 추출액을 세파덱스

LH-20 500ml가 충전된 칼럼에 통과시키고, 메탄올로 용출시켰다. 유효 획분들을 모으고 감압농축시켜 시럽상의 글리옥실일 스페르미딘(II) 디히드로클로라이드 1.583g(수율,33.3%)을 수득하였다.5.55 g (16.26 mmol) of N-seryl-spermidine (I) trihydrochloride was dissolved in 40 ml of water, and 10 ml of an aqueous solution containing 3.55 g (16.58 mmol) of sodium methiiodate was added dropwise at room temperature. . After stirring for 45 minutes, 2N hydrochloric acid was added to the reaction solution to adjust the pH to 1, followed by stirring for 20 minutes, then the pH was adjusted to 4 again by adding 2N NaOH, followed by CM-Sepadex.

500 ml was passed through a packed column. Next, gradient elution was performed using 2.5 l of water and 2.5 l of 1M NaCl solution to collect the effective fractions and dried under reduced pressure. The residual solid obtained is extracted with methanol, and the methanol extract is separated by Sephadex.

500 ml of LH-20 was passed through a packed column and eluted with methanol. The effective fractions were collected and concentrated under reduced pressure to yield 1.583 g (yield, 33.3%) of glyoxyl spermidine (II) dihydrochloride on syrup.

Example 2

1.Synthesis of N- (4-aminobutyl) tartaric acid and monoamide (IX)

13.225 g (150 mmol) of putrescine (VI) is dissolved in 75 ml of tetrahydrofuran (THF), and 10.8 g (50 mmol) of diacetyl-L-tartaric anhydride (VIII) in 450 ml of THF is subjected to ice-cooling stirring. Slowly added dropwise.

After the addition was completed, stirring was continued for 2 hours at room temperature, and after the reaction was completed, the solvent was removed, and then the reaction solution was adjusted to pH 13 by adding 2N NaOH. Next, stirring was carried out at room temperature for 2 hours to carry out hydrolysis of the acetyl group.

1x4(OH^-형) 450ml가 충전된 칼럼에 통과시키고, 물 1.8l로 세척시킨 후 0.5N 아세트산 수용액으로 용출시켰다. 목적 화합물을 함유하는 획분들을 모으고, 감압 건고시켜 백스 결정의 N-(4-아미노부틸)타르타르산 모노아미드(IX) 4.03g(수율,36.6%)을 수득하였다.Dowex the resulting reaction solution

450 ml of 1 × 4 (OH ⁻ type) were passed through a packed column, washed with 1.8 l of water and eluted with a 0.5N acetic acid aqueous solution. Fractions containing the desired compound were collected and dried under reduced pressure to yield 4.03 g (yield, 36.6%) of N- (4-aminobutyl) tartaric acid monoamide (IX) as a white crystal.

NMR (D ₂ O): δ 1.4-1.9 (CH ₂ X ₂ ), 2.98 (CH ₂ NHCO, t, J = 8 Hz), 3.26 (CH ₂ NH, t, J = 7.5 Hz), 4.35 (CHOH, d, J = Hz), 4.49 (CHOH, D, J = 2Hz)

IR (KBr): υ (cm ^-1 ) 3320, 2910, 2850, 1655, 1630, 1550, 1430, 1370, 1310, 1280, 1225, 1130, 1070, 980

2. Synthesis of N- [4- (2-cyanoethyl) aminobutyl] tartaric acid monoamide (XI)

1x4(OH⁺형) 300ml가 충전된 칼럼에 통과시켰다. 다음에 물 1.2l로 세척한 후 0.5N 아세트산수용액으로 용출시켰다. 목적 화합물을 함유하는 획분들을 모으고, 감압농축시켜 시렵상의 N-[4-(2-시아노에틸)아미노부틸]타르타르산 모노아미드(XI) 2.235g (수율, 89.6%)을 수득하였다.In 40 ml of water, 2.01 g (9.127 mmol) of N- (4-aminobutyl) tartaric acid monoamide (IX) obtained in Example 1 was dissolved, 1.273 ml of triethylamine was added, followed by 0.726 ml of acrylonitrile. Was added and the resulting mixture was stirred at rt for 27 h. The reaction solution was then dried under reduced pressure, 20 ml of water was added to the resulting solid, and the resulting solution was added to Dowex.

300 ml of 1 × 4 (OH ⁺ type) were passed through a packed column. It was then washed with 1.2 L of water and eluted with 0.5 N acetic acid solution. Fractions containing the desired compound were collected and concentrated under reduced pressure to yield 2.235 g (yield, 89.6%) of an oxy N- [4- (2-cyanoethyl) aminobutyl] tartaric acid monoamide (XI).

NMR (DMSO-d _S ): δ 1.3-1.8 (CH ₂ X ₂ ), 2.4-3.3 (NCH ₂ , X3, CH ₂ CN), 4.20 (CHOH, X2), 7.7 (CONH)

IR (KBr): υ (cm ^-1 ) 3390, 2930, 2230, 1650, 1630, 1540, 1400, 1120, 1070, 600

3. Synthesis of N- [4- (3-aminopropyl) aminobutyl] tartaric acid monoamide (I)

C-25(Na⁺형) 500ml가 충전된 칼럼에 통과시켰다. 다음에 물 1.5l로 세척시킨 후 물 2l와 1M NaCl수용액 2l를 사용하여 경사용출을 수행하였다.2.101 g (7.688 mmol) of N- [4- (2-cyanoethyl) aminobutyl] tartaric acid monoamide (XI) obtained in Example 2 2 were dissolved in 50 ml of methanol, and 2.195 g of CoCl ₂ , 6H ₂ O was dissolved. (9.226 mmol) and 1.746 g (46.13 mmol) of NaBH ₄ were added slowly with ice-stirring. After completion of the addition, stirring was continued for 3 hours at room temperature to terminate the reaction. Then, 100 ml of water was added, and 2N HCl was added to the reaction solution to adjust the pH to 5.8. Precipitated black crystals were filtered, and the filtrate was concentrated under reduced pressure to remove methanol. 50 ml of water was added to the concentrated solution, and the mixture was diluted with CM-Sepadex.

500 ml of C-25 (Na ⁺ type) were passed through a packed column. Next, the mixture was washed with 1.5 l of water, followed by gradient elution using 2 l of water and 2 l of 1M NaCl solution.

1x4(OH⁺형)500ml가 충전된 칼럼에 통과시킨 다음 물 2l로 세척한후 0.5N 아세트산 수용액으로 용출시켰다. 목적화합물을 함유하는 획분들을 모으고, 감압농축시켜 적색의 시럽상의 N-[4-(3-아미노프로필)아미노부틸]타르타르산 모노아미드(I)·아세테이트 1.225g (수율,39.8%)을 수득하였다.Collect the fractions containing the target compound, Dowox

500 ml of 1 × 4 (OH ⁺ type) was passed through a packed column, washed with 2 liters of water, and eluted with an aqueous 0.5N acetic acid solution. Fractions containing the desired compound were collected and concentrated under reduced pressure to yield 1.225 g (yield, 39.8%) of N- [4- (3-aminopropyl) aminobutyl] tartaric acid monoamide (I) acetate in red syrup. .

NMR (D ₃ O): δ 1.4-2.3 (CH ₂ X 3), 2.8-3.4 (NCH ₂ , X4), 4.25 (CHOH, d, J = 2Hz), 4.4 (CHOH, d, J = 2Hz)

IR (KBr): υ (cm ^-1 ) 3400, 3040, 2950, 1640, 1400, 1120, 1070

4. Synthesis of glyoxyyl spermidine (II)

C-25(Na⁺형) 300ml가 충전된 칼럼에 통과시켰다. 다음에 물 1l로 세척한 후 물 1.5l와 1M NaCl 수용액 1.5l를 사용하여 경상 용출을 수행하였다.1.007 g (2.985 mmol) of N- [4- (3-aminopropyl) aminobutyl] tartaric acid monoamide (I) acetate obtained in Example 3 3 was dissolved in 7 ml of water 80, and metaiodic acid in 20 ml of water. A 650 mg (3.04 mmol) sodium solution was added dropwise with stirring at room temperature. After the dropwise addition, stirring was continued for 30 minutes, and then the reaction solution was added to CM-Sepadex.

300 ml of C-25 (Na ⁺ type) were passed through a packed column. Next, after washing with 1 l of water, light elution was performed using 1.5 l of water and 1.5 l of 1M aqueous NaCl solution.

LH-2001 300ml가 충전된 칼럼에 통과시키고 메탄올로 용출시켰다. 목적화합물을 함유하는 획분들을 모으고, 감압농축시켜 시럽상의 글리옥실일스페르미딘(II)·디히드로클로라이드 681mg(수율,78.1%)을 수득하였다.Fractions containing the desired compound were collected, dried under reduced pressure and extracted with methanol. Sepadex Methanol Extract

300 ml of LH-2001 was passed through a packed column and eluted with methanol. Fractions containing the desired compound were collected and concentrated under reduced pressure to give 681 mg (yield, 78.1%) of glyoxyyl spermidine (II) dihydrochloride on syrup.

Example 3

1.Synthesis of N, N'-bis [4- (2-cyanoethyl) aminobutyl] tartaric acid amide (XIII)

15.45 g (0.075 mole) of diethyl L-tartarate (XII) was added dropwise to 23.31 g (0.165 mole) of cyanoethyl putrescine (V), and after completion of the dropwise addition, the resulting mixture was heated at 80 ° C. for 2 hours. . After cooling the reaction solution, the resulting solids were washed with acetone to obtain 29.64 g of N, N'-bis [4- (2-cyanoethyl) aminobutyl] tartaric acid amide (XIII), a pale yellow crude crystal. Yield, 99.8%).

HP-20(미쓰비시 화학공업사 제품) 1.5l를 함유하는 칼럼에 통과시킨 다음, 물 10l, 5% 메탄올 수용액 5l 및 20% 메탄올 수용액 5l의 순서로 차례로 용출시켰다. 목적 화합물을 함유하는 획분들을 모으고, 감압농축시켜 백색 결정의 N,N'-비스[4-(2-시아노에틸)아미노부틸]타르타르산 아미드(XIII) 3.1g(수율,61.9%)을 수득하였다.5 g of this crystal was dissolved in 20 ml of 0.5 M aqueous NaCl solution, and the resulting solution was equilibrated with 0.5 M aqueous NaCl solution.

The mixture was passed through a column containing 1.5 l of HP-20 (manufactured by Mitsubishi Chemical Corporation), and then eluted in this order in order of 10 l of water, 5 l of 5% aqueous methanol solution and 5 l of 20% aqueous methanol solution. Fractions containing the desired compound were collected and concentrated under reduced pressure to yield 3.1 g (yield, 61.9%) of N, N'-bis [4- (2-cyanoethyl) aminobutyl] tartaric acid amide (XIII) as white crystals. It was.

NMR (DMSO-d _S ): δ 1.3-1.7 (CH ₂ X 4), 2.4-2.9 (CH ₂ N, X6, CH ₂ CN X2), 3.3-3.7 (NH X2, OH X2), 4.20 (CH X2) , 7.60 (CONH X2)

IR (KBr): υ (cm ^-1 ) 3370, 2905, 2220 (CN), 1640, 1520, 1455, 1125

2. Synthesis of N, N'-bis [4- (3-aminopropyl) aminobutyl] tartaric acid amide (I)

C-25(Na⁺형) 500ml가 충전된 칼럼에 통과시켰다. 다음에 물 3l 와 1.5M의 NaCl 수용액 3l를 사용하여 경사용출을 수행하였다. 목적화합물을 함유하는 획분들을 모으고, 감압건조시킨 다음 메탄올로 추출하였다. 메탄올 추출액을 세파덱스

LH-20 500ml가 충전된 칼럼에 통과시키고 메탄올로 용출시켰다. 목적 화합물을 함유하는 획분들을 모으고, 감압건고시켜 시럽상의 N,N'-비스[4-(3-아미노프로필)아미노부틸]타르타르산 아미드(I)·테트라히드로 클로라이드 1.65g(수율, 60.0%)을 수득하였다.19.8 g (50 mmol) of N, N'-bis [4- (2-cyanoethyl) aminobutyl] tartaric acid amide (XIII) crude crystal obtained in Example 3, 1 was dissolved in a mixed solution of 450 ml of methanol and 33 ml of water. 28.55 g (120 mmol) of CoCl ₂ , 6H ₂ O was added followed by the addition of 22.71 g (600 mmol) of NaBH _{4 in} portions under ice-cooling. After the addition of NaBH ₄ , the reaction solution was stirred at room temperature for 1.5 hours, 200 ml of water was added, and 6N hydrochloric acid was added to adjust the pH to 6.5. The black residual solid precipitated was filtered off, and the filtrate was concentrated under reduced pressure to remove methanol. The remaining concentrated solution was diluted to a total volume of 400 ml, and then 40 ml of the resulting solution was prepared by CM-Sepadex.

500 ml of C-25 (Na ⁺ type) were passed through a packed column. Next, gradient elution was performed using 3 l of water and 3 l of a 1.5 M aqueous NaCl solution. Fractions containing the desired compound were collected, dried under reduced pressure and extracted with methanol. Sepadex Methanol Extract

500 ml of LH-20 was passed through a packed column and eluted with methanol. Fractions containing the desired compound were collected and dried under reduced pressure to yield 1.65 g of N, N'-bis [4- (3-aminopropyl) aminobutyl] tartaric acid amide (I) tetrahydrochloride on syrup (yield, 60.0%). Obtained.

NMR (CD ₃ OD): δ 1.5-2.4 (CH ₂ X 6), 2.9-3.4 (NCH ₂ X 8), 4.45 (CH X 2)

IR (KBr): υ (cm ^-1 ) 3380, 2950, 2800, 1640, 1540, 1460, 1125, 1070, 750

3. Synthesis of glyoxyyl spermidine (II)

C-25(Na⁺형) 1000ml가 충전된 칼럼에 통과시켰다. 다음에 물 3l로 세척한 후 물 3l와 0.6M의 NaCl 수용액 3l를 사용하여 경사용출을 행하였다. 목적 화합물을 함유하는 획분들을 모으고, 감압건조시킨 다음 메탄올로 추출하였다. 메탄올 추출액을 세파덱스

LH-20 1000ml가 충전된 칼럼에 통과시키고 메탄올로 용출시켰다. 목적 화합물을 함유하는 획분들을 모으고, 감압농축시켜 시럽상의 글리옥실일스페르미딘(II)·디히드로클로라이드 8.23g(수율,31.3%)을 수득하였다.Aqueous N, N'-bis [4- (3-aminopropyl) aminobutyl] tartaric acid amide (I) was obtained in an aqueous solution containing 16.04 g (75 mmol) of sodium metaperiodate. 360 ml of an aqueous solution containing 45 mmol of tetrahydrochloride was added dropwise and the resulting mixture was stirred at room temperature for 2 hours. After completion of the reaction, 4.5 g (30 mmol) of L-tartaric acid was added thereto, followed by stirring for 15 minutes, and then the yellowish green product precipitated. PH was adjusted to 5.0 by adding 2N NaOH to the filtrate and then CM- Sephadex

1000 ml of C-25 (Na ⁺ type) were passed through a packed column. Next, the mixture was washed with 3 l of water, and then eluted with 3 l of water and 3 l of an aqueous NaCl solution of 0.6 M. Fractions containing the desired compound were collected, dried under reduced pressure and extracted with methanol. Sepadex Methanol Extract

1000 ml of LH-20 was passed through a packed column and eluted with methanol. Fractions containing the desired compound were collected and concentrated under reduced pressure to give 8.23 g (yield, 31.3%) of glyoxyyl spermidine (II) dihydrochloride on syrup.

IR (KBr): υ (cm ^-1 ) 3370, 2950, 1660, 1540, 1460, 1100, 1070

Example 4

C-25(Na⁺형) 12l가 충전된 칼럼에 통과시켰다. 다음에 0.1M 의 NaCl 40l와 0.25M의 NaCl 50l로 단계적으로 용출시켜 목적 화합물을 함유하는 획분들을 모았다. 이들 획분을 물로 4회 희석시키고, CM-세파덱스

C-25(Na⁺형) 7.5l충전된 칼럼에 통과시킨 다음 0.8M의 NaCl로 용출시켰다. 목적화합물을 함유하는 획분을 모으고 동결건조시켰다. 동결건조 생성물을 메탄올 4l로 추출하여 메탄올 추출액을 감압건고시켜 시럽상의 글리옥실일스페르미딘(II)·디히드로클로라이드 400.4g(디에틸타르타레이트(XIII)로부터의 총 수율,43.5%)을 수득하였다.To 450 g (3.19 mol) of cyanoputresin (V) was added 300 g (1.45 mmol) of diethyl L-tartarate (XII), and the resulting mixture was heated at 80 ° C. for 3 hours. The reaction mixture was dissolved in 4 l of methanol, and saturated with ammonia gas, while cooling the resulting solution. Then, 180 g of Raney-Nickel was added, and hydrogenation was performed at 40 ° C. under a hydrogen pressure of 12 kg / cm ² for 24 hours. After completion of the reaction, the catalyst was filtered off and the filtrate was concentrated under reduced pressure. The syrup material thus obtained was dissolved in 3 l of water, and 6N hydrochloric acid was added thereto to adjust the pH to 5.0. A solution of 397 g (1.66 mol) of sodium metaperiodate in 5 l of water was then added in portions to the solution and the reaction mixture was stirred at room temperature for 3 hours. After the reaction was completed, 10 ml of ethylene glycol was added to the reaction solution to react all remaining reaction reagents. 6N NaOH was then added to this solution to adjust the pH to 5.0, diluted four times with water, and then CM-Sepadex.

12 liters of C-25 (Na ⁺ type) were passed through a packed column. Next, the fractions containing the target compound were collected by eluting stepwise with 40 L of 0.1 M NaCl and 50 L of 0.25 M NaCl. These fractions were diluted four times with water and CM- Sephadex

C-25 (Na ⁺ type) was passed through a 7.5 L packed column and eluted with 0.8 M NaCl. Fractions containing the desired compound were collected and lyophilized. The lyophilized product was extracted with 4 L of methanol and the methanol extract was dried under reduced pressure to give 400.4 g of glyoxyylspermidine (II) dihydrochloride on syrup (total yield from diethyl tartrate (XIII), 43.5%). It was.

Example 5

Synthesis of N- [4- (3-aminopropyl) aminobutyl] -2- (7-guanidinoheptanamido) -2-hydroxyethanamide

C-25(Na 형), 150ml가 충전된 칼럼(내부 직경 20mm)에 통과시키고, 물 1.5l와 0.8M의 염화나트륨 수용액 1.5l를 사용하여 경사용출에 의해 분획시켰다. 목적 화합물을 함유하는 획분들을 모으고, 농축시킨 다음 메탄올 10ml로 3회 추출하였다. 메탄올층을 세파덱스

LH-20 150ml가 충전된 칼럼에 통과시키고, 메탄올로 전개시켰다. 목적 화합물을 함유하는 획분들을 모으고, 증발건고시켜 백색분말상의 N-[4-(3-아미노프로필)아미노부틸]-2-(7-구아니노헵탄아미도)-2-히드록시에탄아미드·트리히드로클로라이드 317mg(수율,39%)을 수득하였다.360 mg (1.62 mmol) of 7-guanidinoheptanamido hydrochloride, N- [4- (3-aminopropyl) aminobutyl] -2,2-dihydroxyethaneamide dihydrochloride obtained in Example 4 A mixture of 568 mg (1.94 mmol), 214 mg glutamic acid (1.62 mmol), and 0.36 ml of water was heated at 60 ° C. for 24 hours. After the reaction was completed, 5 ml of water was added to the reaction mixture, followed by CM-Sepadex.

C-25 (Na type), passed through a column packed with 150 ml (inner diameter 20 mm), and fractionated by decanting using 1.5 l of water and 1.5 l of 0.8 M aqueous sodium chloride solution. Fractions containing the desired compound were collected, concentrated and extracted three times with 10 ml of methanol. Methanol Sephadex

150 ml of LH-20 was passed through a packed column and developed with methanol. Fractions containing the desired compound were collected, evaporated to dryness, and then N- [4- (3-aminopropyl) aminobutyl] -2- (7-guaninoheptanamido) -2-hydroxyethanamide on white powder. 317 mg (39%) of trihydrochloride were obtained.

Example 6

Synthesis of N- [4- (3-aminopropyl) aminobutyl] -2- (7-guaninoheptanamido) -2-hydroxyethanamide

18 g (80.9 mmol) of 7-guanidinoheptanamido hydrochloride, N- [4- (3-aminopropyl) aminobutyl] -2,2-dihydroxyethaneamidedihydrochloride 23.6 obtained in Example 4 A mixture of g (80.9 mmol), 5.7 g (27 mmol) citric acid, and 200 ml of water was evaporated to dryness to obtain a syrup material containing 1.6 g of water. The syrup material was heated at 60 ° C. for 8 hours and purified in a similar manner as in Example 5 to give N- [4- (3-aminopropyl) aminobutyl] -2- (7-guaninoheptane in white powder form. Amido) -2-hydroxyethaneamidetrihydroxychloride 19.0g (yield, 47.3%) was obtained.

Example 7

Synthesis of N- [4- (3-aminopropyl) aminobutyl] -2- (9-guanidinoamide) -2-hydroxyethanamide

C-25(Na^-형)과 세파덱스

LH-20을 사용하여 실시예 5에서와 유사한 방법으로 혼합물의 정제를 행하여 백색 분말상의 N-[4-(3-아미노프로필)아미노부틸]-2-(9-구아니디노노난아미드)-2-히드록시에탄아미드·트리히드로클로라이드 324mg (수율, 49%)을 수득하였다.316 mg (1.26 mmol) of 9-guanidinononanamide hydrochloride, N- [4- (3-aminopropyl) aminobutyl] -2,2-dihydroxyethanamide dihydrochloride 442 mg obtained in Example 4 (1.51 mmol), 166 mg (1.26 mmol) glutaric acid, and 0.01 ml of water were heated at 60 ° C. for 24 hours. CM- Sephadex after completion of reaction

C-25 (Na^-Type) and Sephadex

Purification of the mixture in a similar manner as in Example 5 using LH-20, yielding white powdery N- [4- (3-aminopropyl) aminobutyl] -2- (9-guanidinononanamide) -2. 324 mg (yield, 49%) of hydroxyethanamide trihydrochloride were obtained.

Claims (5)

N- [4-3-aminopropyl) aminobutyl] -2,2-dihydrate represented by the following structural formula (II), characterized by oxidative cleavage of the CC bond of the compound represented by the following general formula (I) Process for the preparation of oxyethanamide:

In the above formula, R ¹ is a hydroxy group or an amino group, but two, R ¹ cannot be an amino group at the same time, and R ¹ is a hydrogen atom, a carboxyl group, or CONH (CH ₂ ) ₄ NH ₂ (CH ₂ ) ₃ NH ₂ . The method for producing a compound of formula (II) according to claim 1, wherein the compound represented by formula (I) is a compound of the following structural formula.

The method for producing a compound of formula (II) according to claim 1, wherein the compound represented by formula (I) is a compound of the following structural formula.

The method for producing a compound of formula (II) according to claim 1, wherein the compound represented by formula (I) is a compound of the following structural formula.

After oxidative cleavage of the CC bond of the compound represented by the following general formula (I) to prepare N- [4- (3-aminopropyl) aminobutyl] -2,2-dihydroxy ethaneamide of the following structural formula (II) A process for preparing a mixture of formula (XV), consisting of condensing ethanamide of formula (II) prepared above with W-guanidino fatty acid amide of formula (XVI):

In the above formulas, R ¹ is a hydroxy group or an amino group, but two R ¹ may not be an amino group at the same time, and R ² is a hydrogen atom, a carboxyl group, or CONH (CH ₂ ) ₄ NH (CH | ₂ ) ₃ NH ₂ and n is an integer of 6-8.